Generate functional, human-specific sensory neurons in a dish with this easy-to-use culture system. STEMdiff™ Sensory Neuron Maturation Kit is the final step in a workflow beginning from human pluripotent stem cells (hPSCs) and leading to a population of sensory neurons that can be used for drug discovery and pain research applications. After obtaining PSC-derived neural crest cells (NCCs) with STEMdiff™ Neural Crest Differentiation Kit, use the serum-free STEMdiff™ Sensory Neuron Differentiation Kit to generate sensory neuron precursors. STEMdiff™ Sensory Neuron Maturation Kit can then be used to maintain and mature the sensory neuron precursors. The resulting cells are BRN3A-positive and more than 70% TUJ1-positive. With the physiological glucose and osmolarity conditions provided by BrainPhys™ as the basal medium, the neurons exhibit activity in response to sensory ligands and changes in temperature.
Area of Interest Disease Modeling, Drug Discovery and Toxicity Testing, Neuroscience
Formulation Category Serum-Free
Data Figures
Figure 1. Schematic for the STEMdiff™ Sensory Neuron Culture System Protocol
Sensory neurons can be generated in 7 days from sensory neuron precursors after 6 days in STEMdiff™ Sensory Neuron Differentiation Medium. For the differentiation of sensory neuron precursors from hPSC-derived neural crest cells, see the PIS.
Figure 2. STEMdiff™ Sensory Neuron Kits Promote Differentiation Across Multiple Embryonic Stem and Induced Pluripotent Stem Cell Lines
NCCs generated from hPSCs in mTeSR™ Plus using the STEMdiff™ Neural Crest Differentiation Kit were differentiated and matured to sensory neurons using the STEMdiff™ Sensory Neuron Differentiation and Maturation Kits. (A) Sensory neurons were generated after hPSC-derived NCCs were cultured with the STEMdiff™ Sensory Neuron Differentiation Kit for 6 days and then the STEMdiff™ Sensory Neuron Maturation Kit for 6 days. The resulting cultures contain a population of cells expressing sensory neuron markers peripherin (green) and BRN3A (red) along with (B) neuronal marker class III β-tubulin (TUJ1, red). (C) Midbrain neuron controls generated with STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits do not have detectable peripherin (green) or BRN3A (red) expression, although they express (D) neuronal marker class III β-tubulin (TUJ1, red). Nuclei are labeled with DAPI (blue). Human ES and iPS cell lines were maintained in either mTeSR™1, TeSR™-E8™, or mTeSR™ Plus and differentiated with STEMdiff™ Neural Crest Differentiation Kit, followed by STEMdiff™ Sensory Neuron Differentiation and Maturation Kits. The percentage expression of (E) BRN3A+ and (F) TUJ1+ cells in the resulting cultures was quantified. This differentiation generated BRN3A+ sensory neurons (25.3% ± 6.9%, mean ± SEM; n=7 cell lines, 3 - 23 replicates per condition) that expressed neuronal marker class III β-tubulin (TUJ1; 90.3% ± 4.1%, mean ± SEM; n=4 cell lines, 3 - 12 replicates per condition). Numbers are % positive over total DAPI in a tiled image. NCCs = neural crest cells; hPSCs = human pluripotent stem cells; ES = embryonic stem; iPS = induced pluripotent stem
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